Multicenter evaluation of the Vetscan Imagyst system using Ocus 40 and EasyScan One scanners to detect gastrointestinal parasites in feces of dogs and cats.

The Vetscan Imagyst system (Zoetis) is a novel, artificial intelligence-driven detection tool that can assist veterinarians in the identification of enteric parasites in dogs and cats. This system consists of a sample preparation device, an automated digital microscope scanner, and a deep-learning algorithm. The EasyScan One scanner (Motic) has had good diagnostic performance compared with manual examinations by experts; however, there are drawbacks when used in veterinary practices in which space for equipment is often limited. To improve the usability of this system, we evaluated an additional scanner, the Ocus 40 (Grundium). Our objectives were to 1) qualitatively evaluate the performance of the Vetscan Imagyst system with the Ocus 40 scanner for identifying Ancylostoma, Toxocara, and Trichuris eggs, Cystoisospora oocysts, and Giardia cysts in canine and feline fecal samples, and 2) expand the assessment of the performance of the Vetscan Imagyst system paired with either the Ocus 40 or EasyScan One scanner to include a larger dataset of 2,191 fecal samples obtained from 4 geographic regions of the United States. When tested with 852 canine and feline fecal samples collected from different geographic regions, the performance of the Vetscan Imagyst system combined with the Ocus 40 scanner was correlated closely with manual evaluations by experts. Sensitivities were 80.0‒97.0% and specificities were 93.7‒100.0% across the targeted parasites. When tested with 1,339 fecal samples, the Vetscan Imagyst system paired with the EasyScan One scanner successfully identified the targeted parasite stages; sensitivities were 73.6‒96.4% and specificities were 79.7‒100.0%.

Seasonality of Amblyomma americanum (Acari: Ixodidae) Activity and Prevalence of Infection with Tick-Borne Disease Agents in North Central Oklahoma.

Background: Amblyomma americanum is the most common tick infesting both animals and humans in the southern United States and transmits a variety of zoonotic agents. The rise in tick-borne diseases (TBD) globally imparts a need for more active surveillance of tick populations to accurately quantify prevalence and risk of tick-borne infectious organisms. To better understand TBD risk in north central Oklahoma, this study aimed to describe the current seasonal activity of A. americanum in this region and investigate the seasonality of tick-borne infectious agents. Materials and Methods: Tick collections were performed twice a month for a duration of 2 years at a field site in Payne County, Oklahoma. Total nucleic acid was extracted from a subset of adult A. americanum and tested for Rickettsia spp., Ehrlichia spp., and Borrelia spp. using established PCR protocols. Results: Peak activity times for each life stage were observed, with adults primarily active 1 month earlier than historical seasonal trends describe, and male A. americanum active earlier in the year than female A. americanum. Rickettsia spp., Ehrlichia chaffeensis, Ehrlichia ewingii, and Borrelia lonestari were found in 26.4%, 6.1%, 2.5%, and 1.1% of adult A. americanum, respectively. No seasonal trend in spotted fever group Rickettsia spp. (SFGR) was observed in peak activity months. Conclusions: This study found an apparently shifting phenology for A. americanum adults in Oklahoma. While these results did not show a trend in SFGR, further investigation is needed to better understand the potential seasonality of infection prevalence within A. americanum across the expanding range of this vector, especially considering the extended activity of males in winter months.

Fluralaner (Bravecto®) treatment kills Aedes aegypti after feeding on Dirofilaria immitis-infected dogs.

BACKGROUND: Transmission of canine heartworm (Dirofilaria immitis) from infected to naïve dogs is dependent on successful mosquito feeding and survival. METHODS: To determine whether treating heartworm-infected dogs with fluralaner (Bravecto®) limits the survival of infected mosquitoes, and potentially the transmission of D. immitis, we allowed female mosquitoes to feed on microfilaremic dogs and evaluated mosquito survival and infection with D. immitis. Eight dogs were experimentally infected with D. immitis. On day 0 (~ 11 months post-infection), four microfilaremic dogs were treated with fluralaner according to label directions while the other four were non-treated controls. Mosquitoes (Aedes aegypti Liverpool) were allowed to feed on each dog on days -7, 2, 30, 56, and 84. Fed mosquitoes were collected, and the number of live mosquitoes determined at 6 h, 24 h, 48 h, and 72 h post-feeding. Surviving mosquitoes held for 2 weeks were dissected to confirm third-stage D. immitis larvae; PCR (12S rRNA gene) was performed post-dissection to identify D. immitis in mosquitoes. RESULTS: Prior to treatment, 98.4%, 85.1%, 60.7%, and 40.3% of mosquitoes fed on microfilaremic dogs were alive at 6 h, 24 h, 48 h, and 72 h post-feeding, respectively. Similarly, mosquitoes fed on microfilaremic, non-treated dogs were alive 6 h post-feeding (98.5-100%) throughout the study. In contrast, mosquitoes fed on fluralaner-treated dogs 2 days after treatment were dead or severely moribund by 6 h post-feeding. At 30 and 56 days post-treatment, > 99% of mosquitoes fed on treated dogs were dead by 24 h. At 84 days post-treatment, 98.4% of mosquitoes fed on treated dogs were dead by 24 h. Before treatment, third-stage larvae of D. immitis were recovered from 15.5% of Ae. aegypti 2 weeks after feeding, and 72.4% were positive for D. immitis by PCR. Similarly, 17.7% of mosquitoes fed on non-treated dogs had D. immitis third-stage larvae 2 weeks after feeding, and 88.2% were positive by PCR. Five mosquitoes fed on fluralaner-treated dogs survived 2 weeks post-feeding, and 4/5 were from day 84. None had third-stage larvae at dissection, and all were PCR-negative. CONCLUSION: The data indicate that fluralaner treatment of dogs kills mosquitoes and thus would be expected to reduce transmission of heartworm in the surrounding community.

Safety and effectiveness of the sodium-glucose cotransporter inhibitor bexagliflozin in cats newly diagnosed with diabetes mellitus.

BACKGROUND: Bexagliflozin is a sodium-glucose cotransporter 2 (SGLT2) inhibitor. A pilot study has shown that bexagliflozin can decrease dependence on exogenous insulin in cats with diabetes mellitus (DM). OBJECTIVE: To evaluate the safety and effectiveness of bexagliflozin as a monotherapy for DM in previously untreated cats. ANIMALS: Eighty-four client-owned cats. METHODS: Historically controlled prospective open-label clinical trial. Cats were dosed PO with 15 mg bexagliflozin once daily for 56 days, with a 124-day extension to evaluate safety and treatment effect durability. The primary endpoint was the proportion of cats experiencing a decrease in hyperglycemia and improvement in clinical signs of hyperglycemia from baseline on day 56. RESULTS: Of 84 enrolled cats, 81 were evaluable on day 56, and 68 (84.0%) were treatment successes. Decreases in mean serum glucose, fructosamine, and ß-hydroxybutyrate (ß-OHB) concentrations were observed, and investigator assessments of cat neurological status, musculature, and hair coat quality improved. Owner evaluations of both cat and owner quality of life were favorable. The fructosamine half-life in diabetic cats was found to be 6.8 days. Commonly observed adverse events included emesis, diarrhea, anorexia, lethargy, and dehydration. Eight cats experienced serious adverse events, 3 of which led to death or euthanasia. The most important adverse event was euglycemic diabetic ketoacidosis, diagnosed in 3 cats and presumed present in a fourth. CONCLUSION AND CLINICAL IMPORTANCE: Bexagliflozin decreased hyperglycemia and observed clinical signs in cats newly diagnosed with DM. As a once-daily PO medication, bexagliflozin may simplify management of DM in cats.

Geographic Distribution and Seasonality of Brown Dog Tick Lineages in the United States.

Two lineages of brown dog ticks (Rhipicephalus sanguineus sensu lato (s.l.), Latreille [Acari: Ixodidae]) have been described in North America: temperate and tropical. To characterize the distribution of these lineages across this region and evaluate seasonal activity, a 12S rRNA mitochondrial gene fragment was sequenced from R. sanguineus s.l. collected from hundreds of dogs and cats from different locations across 25 of the 50 states from 2018 to 2021. Infestations with temperate lineage predominated (78.5%) and were identified on pets from 20 states, with most (83.5%) from areas with annual mean daily average temperature <20°C. Tropical lineage submissions were less common (19.3%), submitted from 15 states, and most (80.0%) tropical lineage ticks were from areas with an annual mean daily average temperature >20°C. Although travel history was not obtained for all dogs, when tropical lineage infestations were found in colder regions, follow up conversations with veterinarians suggested some of these infestations may have resulted from recent travel of dogs. A limited number (2.2%) of dogs from Arizona and Texas were co-infested with both lineages. Both temperate and tropical lineage ticks were collected from pets in every month of the year. Temperate lineage infestations were primarily collected March through August while tropical lineage infestations were more often collected June through November. These data confirm at least two lineages of R. sanguineus s.l. are present in the United States, each predominating in distinct, overlapping geographies, and suggest that peak activity of each lineage occurs at different times of the year.

Diversity and geographic distribution of rickettsial agents identified in brown dog ticks from across the United States.

Rhipicephalus sanguineus sensu lato, or brown dog ticks, transmit a variety of pathogens of veterinary and public health importance globally. Pathogens vectored by brown dog ticks and identified in the United States include Babesia vogeli, Ehrlichia canis, and several spotted fever group Rickettsia spp. (SFGR). Due to the challenge of collecting canine blood samples nationwide to screen for exposure to these pathogens, we took an indirect approach and tested brown dog ticks for molecular evidence of infection. Brown dog ticks (616 adults and 65 nymphs) collected from dogs and cats across the nation were tested by separate PCR assays detecting Babesia spp., E. canis, and SFGR. While no Babesia sp. was found, we identified rickettsial agents in 3.5% (24/681; 95% CI 2.4-5.2%) of the ticks. Pathogens and related organisms detected in ticks included E. canis (n = 1), Rickettsia amblyommatis (n = 3), Rickettsia massiliae (n = 11), Rickettsia monacensis (n = 3), Rickettsia montanensis (n = 5), and an undefined Rickettsia species (n = 1). These data demonstrate a wider geographic distribution of R. massiliae than previously known, and to the authors' knowledge, reports R. monacensis in brown dog ticks for the first time. Due to the close association that brown dog ticks have with domestic dogs and humans, more research is needed to understand the full array of organisms, some of which are zoonotic, potentially transmitted by this widespread tick complex.

Survey on the Presence of Equine Tick-Borne Rickettsial Infections in Southcentral United States.

Although ticks are known vectors of pathogens across a range of hosts, there is limited research on emerging tick-borne diseases of horses in the United States. Tick surveys from other regions suggest Rickettsia spp. and Ehrlichia spp. may be clinically relevant in horses. To better understand the transmission risk of these tick-borne rickettsial disease agents to horses, ticks were collected from horses in Oklahoma. Ticks for the current study (306 Amblyomma americanum, 20 Dermacentor albipictus, 19 D. variabilis, and 7 A. maculatum) were evaluated for Rickettsia spp. and Ehrlichia spp. using polymerase chain reaction and sequence analysis. Serum samples from infested and noninfested horses were evaluated for antibodies to R. rickettsii using indirect fluorescence antibody testing and Ehrlichia spp. and Anaplasma spp. using a commercial enzyme-linked immunoassay. Of the horses with tick infestations, 71.4% hosted at least one tick with a rickettsial agent detected. Rickettsia spp. were identified in 25.9% (91/352) of the ticks tested with R. amblyommatis (80.2%; 73/91) most often detected. Ehrlichia spp. were identified in 2.8% (10/352) of the ticks tested with E. ewingii most often identified. Serologic screening revealed no horses with antibodies to R. rickettsii or Anaplasma spp., but 29.6% of the examined horses had circulating antibodies to Ehrlichia spp. Together, these results demonstrate the presence of Rickettsia spp. and Ehrlichia spp. in equine ticks and evidence of past or current infection with Ehrlichia spp. in Oklahoma horses which strongly suggests there is a need to explore the relationship between these agents and equine health.

Influence of tick sex and geographic region on the microbiome of Dermacentor variabilis collected from dogs and cats across the United States.

As tick-borne diseases continue to increase across North America, current research strives to understand how the tick microbiome may affect pathogen acquisition, maintenance, and transmission. Prior high throughput amplicon-based microbial diversity surveys of the widespread tick Dermacentor variabilis have suggested that life stage, sex, and geographic region may influence the composition of the tick microbiome. Here, adult D. variabilis ticks (n = 145) were collected from dogs and cats from 32 states with specimens originating from all four regions of the United States (West, Midwest, South, and Northeast), and the tick microbiome was examined via V4-16S rRNA gene amplification and Illumina sequencing. A total of 481,246 bacterial sequences were obtained (median 2924 per sample, range 399-11,990). Fifty genera represented the majority (>80%) of the sequences detected, with the genera Allofrancisella and Francisella being the most abundant. Further, 97%, 23%, and 5.5% of the ticks contained sequences belonging to Francisella spp., Rickettsia spp., and Coxiella spp., respectively. No Ehrlichia spp. or Anaplasma spp. were identified. Co-occurrence analysis, by way of correlation coefficients, between the top 50 most abundant genera demonstrated five strong positive and no strong negative correlation relationships. Geographic region had a consistent effect on species richness with ticks from the Northeast having a significantly greater level of richness. Alpha diversity patterns were dependent on tick sex, with males exhibiting higher levels of diversity, and geographical region, with higher level of diversity observed in ticks obtained from the Northeast, but not on tick host. Community structure, or beta diversity, of tick microbiome was impacted by tick sex and geographic location, with microbiomes of ticks from the western US exhibiting a distinct community structure when compared to those from the other three regions (Northeast, South, and Midwest). In total, LEfSe (Linear discriminant analysis Effect Size) identified 18 specific genera driving these observed patterns of diversity and community structure. Collectively, these findings highlight the differences in bacterial diversity of D. variabilis across the US and supports the interpretation that tick sex and geographic region affects microbiome composition across a broad sampling distribution.

Identification of Rickettsia spp. and Babesia conradae in Dermacentor spp. Collected from Dogs and Cats Across the United States.

In the United States, Dermacentor variabilis and Dermacentor andersoni are considered key vectors for Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever. Through regional surveillance, a wide diversity of Rickettsia spp. have been documented in D. variabilis, and Dermacentor spp. has been suggested as potential vectors for various other pathogens, including Babesia spp. and Ehrlichia canis. To better define the prevalence and diversity of pathogens in Dermacentor spp. across the United States, 848 ticks collected from dogs and cats in 44/50 states in 2018-2019 were tested by PCR for Rickettsia spp.-specific 17 kDa and ompA gene fragments; a subset of Dermacentor spp. was also tested with PCR, targeting fragments of the 18S and large subunit region rRNA genes of Babesia spp. and 16S rRNA genes of E. canis. Rickettsia spp. was identified in 12.5% (106/848) of ticks. Species detected include Rickettsia montanensis (n = 64 ticks), Rickettsia bellii (n = 15 ticks), Rickettsia rhipicephali (n = 13 ticks), Rickettsia peacockii (n = 8 ticks), Rickettsia amblyommatis (n = 3 ticks), Rickettsia cooleyi (n = 1 tick), and unclassified Rickettsia spp. (n = 2 ticks). Ticks with R. montanensis and R. bellii were submitted from every U.S. region; R. rhipicephali was predominantly detected in ticks from the southern half of the United States, and all R. peacockii-positive ticks were D. andersoni that originated from the Rocky Mountain states. Ehrlichia canis was not detected in any Dermacentor spp., and Babesia conradae was detected in two Dermacentor albipictus. Because most ticks had fed on dogs or cats before submission, these findings do not implicate a given Dermacentor sp. as a primary vector of these agents, but in regard to Rickettsia spp., the data do support other published work showing D. variabilis harbors a diversity of Rickettsia species with unknown implications for animal and human health.

Molecular detection of Cercopithifilaria bainae in brown dog ticks collected from dogs across the United States.

Cercopithifilaria bainae is a filarioid nematode of dogs shown to use Rhipicephalus sanguineus sensu lato (s.l.), the brown dog tick, as the vector. Previously in the United States, C. bainae infections have been reported in a dog from Florida, and in dogs and ticks in Oklahoma, but data are lacking from other areas of the country. Here, we tested brown dog ticks from across the United States for C. bainae DNA to assess the geographic distribution of where this novel parasite may be cycling in ticks and dogs. Archival brown dog ticks were available for testing through the national tick survey Show Us Your Ticks. Ticks were morphologically identified, dissected, and tested by PCR to detect filarioid mitochondrial DNA. A total of 1400 brown dog ticks were tested from 321 separate animals from 23 states, with 5.7 % (80/1400) of the ticks testing positive for C. bainae DNA. At least one positive tick was detected in submissions from 9 states in addition to Florida and Oklahoma. Cercopithifilaria bainae DNA was detected in larval, nymphal, and adult stages of brown dog ticks and only in ticks removed from dogs. Of all dogs with brown dog ticks collected from them, 17.6 % (55/312) were infested with at least one tick that harbored C. bainae DNA. Findings from this study demonstrate a wider geographic range of C. bainae than previously known, and that dogs are commonly infested with brown dog ticks with molecular evidence of C. bainae infection.

Equine attachment site preferences and seasonality of common North American ticks: Amblyomma americanum, Dermacentor albipictus, and Ixodes scapularis.

BACKGROUND: Ticks are common on horses, but recent publications characterizing equine tick infestations in North America are lacking. METHODS: To further understand attachment site preferences of common ticks of horses, and to document the seasonality of equine tick infestation in northeastern Oklahoma, horses from eight farms were evaluated twice a month over a 1-year period. Each horse was systematically inspected beginning at the head and moving caudally to the tail. Attachment sites of ticks were recorded and all ticks collected were identified to species and stage. RESULTS: Horses (26 males and 62 females) enrolled in the study ranged in age from 1 to 23 years (mean = 12, 95% CI 11-13). A total of 2731 ticks were collected; 84.1% (74/88) of the horses were infested (median = 3 ticks) at one or more examinations. Five tick species were identified, including Amblyomma americanum (78.2%; 2136/2731), Ixodes scapularis (18.2%; 497/2731), Dermacentor albipictus brown variant (2.6%; 71/2731), Dermacentor variabilis (0.7%; 20/2731), and Amblyomma maculatum (0.3%; 7/231). Most ticks were adults (83.6%; 2282/2731), but immature A. americanum (436/2136; 20.4%), D. albipictus (12/71; 16.9%), and A. maculatum (n = 1) were occasionally recovered. Amblyomma americanum were most often attached to the inguinal area, and I. scapularis and D. albipictus were most commonly found on the chest and axillary region (P < 0.0001). Ticks were found on horses in every month of the year. The largest number of ticks (638/2731; 23.4%) were collected in May (P < 0.0001). Amblyomma americanum, primarily immature, was the only tick recovered in September, I. scapularis and D. albipictus predominated October through February, and both A. americanum and I. scapularis were common in March. In the warmer months, April through August, A. americanum was the most common tick, followed by D. variabilis and A. maculatum. CONCLUSIONS: This research confirms that ticks common on horses in North America have attachment site preferences and that ticks infest horses in Oklahoma throughout the year, including during the winter. Additional research is warranted to fully understand the risk these infestations pose to equine health.

Ticks infesting dogs and cats in North America: Biology, geographic distribution, and pathogen transmission.

A diverse array of ixodid and argasid ticks infest dogs and cats in North America, resulting in skin lesions, blood loss, and disease. The ticks most commonly found on pets in this region are hard ticks of the genera Amblyomma, Dermacentor, Ixodes, and Rhipicephalus, as well as the more recently established Haemaphysalis longicornis. Soft tick genera, especially Otobius and Ornithodoros, are also reported from pets in some regions. In this review, we provide a summary of the complex and diverse life histories, distinct morphologies, and questing and feeding behaviors of the more common ticks of dogs and cats in North America with a focus on recent changes in geographic distribution. We also review pathogens of dogs and cats associated with the different tick species, some of which can cause serious, potentially fatal disease, and describe the zoonotic risk posed by ticks of pets. Understanding the natural history of ticks and the maintenance cycles responsible for providing an ongoing source of tick-borne infections is critical to effectively combatting the challenges ticks pose to the health of pets and people.

Tick salivary gland extract induces alpha-gal syndrome in alpha-gal deficient mice.

INTRODUCTION: Alpha-gal syndrome (AGS) is characterized by delayed hypersensitivity to non-primate mammalian meat in people having specific immunoglobulin E (sIgE) to the oligosaccharide galactose-alpha-1,3-galactose. AGS has been linked to tick bites from Amblyomma americanum (Aa) in the U.S. A small animal model of meat allergy is needed to study the mechanism of alpha-gal sensitization, the effector phase leading to delayed allergic responses and potential therapeutics to treat AGS. METHODS: Eight- to ten-weeks old mice with a targeted inactivation of alpha-1,3-galactosyltransferase (AGKO) were injected intradermally with 50 µg of Aa tick salivary gland extract (TSGE) on days 0, 7, 21, 28, 42, and 49. Total IgE and alpha-gal sIgE were quantitated on Day 56 by enzyme-linked immunosorbent assay. Mice were challenged orally with 400 mg of cooked pork kidney homogenate or pork fat. Reaction severity was assessed by measuring a drop in core body temperature and scoring allergic signs. RESULTS: Compared to control animals, mice treated with TSGE had 190-fold higher total IgE on Day 56 (0.60 ± 0.12 ng/ml vs. 113.2 ± 24.77 ng/ml; p < 0.001). Alpha-gal sIgE was also produced in AGKO mice following TSGE sensitization (undetected vs. 158.4 ± 72.43 pg/ml). Further, sensitized mice displayed moderate clinical allergic signs along with a drop in core body temperature of ≥2°C as an objective measure of a systemic allergic reaction. Interestingly, female mice had higher total IgE responses to TSGE treatment but male mice had larger declines in mean body temperature. CONCLUSION: TSGE-sensitized AGKO mice generate sIgE to alpha-gal and demonstrate characteristic allergic responses to pork fat and pork kidney. In keeping with the AGS responses documented in humans, mice reacted more rapidly to organ meat than to high fat pork challenge. This mouse model establishes the central role of tick bites in the development of AGS and provides a small animal model to mechanistically study mammalian meat allergy.

Optimizing heartworm diagnosis in dogs using multiple test combinations.

BACKGROUND: Various heartworm (HW) diagnostic testing modalities detect products of, or reactions to, different life cycle stages of Dirofilaria immitis. Microfilariae (Mf) can be directly visualized in blood, antigen (Ag) from immature and adult heartworms may be detected on commercial assays, and antibody (Ab) tests detect the host immune response to larval stages. Ag and Mf tests are commonly used in dogs, which frequently carry adult HW infections, but Ab tests have only been validated for use in cats. In some HW-infected dogs, Ag is blocked by immune complexing leading to false-negative results. Heat-treatment (HT) to disrupt these complexes can increase the sensitivity of HW Ag tests. The aim of this study was to compare different methods for diagnosing HW infection in dogs at high risk using individual and paired diagnostic tests, including an exploration of using Ab tests designed for cats to test canine samples. METHODS: One hundred stray adult (≥ 2-year-old) dogs in Florida shelters were tested using Mf, HW Ag, and HW Ab tests (feline HW Ab tests currently not commercially validated/approved for use in dogs); two versions of each test platform were used. RESULTS: Fourteen dogs tested positive using point-of-care (POC) Ag tests; an additional 2 dogs tested positive with microtiter well assay, and an additional 12 dogs tested positive using HT Ag testing. For individual tests, Ag test sensitivity/specificity compared to HT Ag was 50-57%/100%, and Ab tests were 46-64%/82-94%. Sensitivity estimates for individual tests were higher when comparing to non-HT Ag. Pairing POC Ag tests with Mf tests improved sensitivity without loss of specificity, while pairing POC Ag and Ab tests modestly increased sensitivity at the expense of specificity. CONCLUSIONS: Screening dogs for HW infection using both POC Ag and Mf detection, which is recommended by the American Heartworm Society, improved diagnostic performance in this study compared to single Ag test use, but may have missed more than one in four infected dogs. The need to improve access to highly accurate, rapid, and inexpensive large-scale HW testing for dogs in animal shelters remains largely unmet by current testing availability. The development of practical and validated protocols that incorporate heat or chemical treatment to disrupt Ag-Ab complexes in POC testing or decreasing the cost and time required for such testing in reference laboratories might provide solutions to this unmet need. Similar studies performed in countries where the prevalence of parasites such as D. repens or A. vasorum is different to the USA could potentially yield very different positive predictive values for both HT and non-HT Ag tests.

Dermacentor variabilis is the Predominant Dermacentor spp. (Acari: Ixodidae) Feeding on Dogs and Cats Throughout the United States.

Throughout North America, Dermacentor spp. ticks are often found feeding on animals and humans, and are known to transmit pathogens, including the Rocky Mountain spotted fever agent. To better define the identity and distribution of Dermacentor spp. removed from dogs and cats in the United States, ticks submitted from 1,457 dogs (n = 2,924 ticks) and 137 cats (n = 209 ticks) from veterinary practices in 44/50 states from February 2018-January 2020 were identified morphologically (n = 3,133); the identity of ticks from regions where Dermacentor andersoni (Stiles) have been reported, and a subset of ticks from other regions, were confirmed molecularly through amplification and sequencing of the ITS2 region and a 16S rRNA gene fragment. Of the ticks submitted, 99.3% (3,112/3,133) were Dermacentor variabilis (Say), 0.4% (12/3,133) were D. andersoni, and 0.3% (9/3,133) were Dermacentor albipictus (Packard). While translocation of pets prior to tick removal cannot be discounted, the majority (106/122; 87%) of Dermacentor spp. ticks removed from dogs and cats in six Rocky Mountain states (Montana, Idaho, Wyoming, Nevada, Utah, and Colorado) were D. variabilis, suggesting this species may be more widespread in the western United States than is currently recognized, or that D. andersoni, if still common in the region, preferentially feeds on hosts other than dogs and cats. Together, these data support the interpretation that D. variabilis is the predominant Dermacentor species found on pets throughout the United States, a finding that may reflect recent shifts in tick distribution.

Ixodes spp. from Dogs and Cats in the United States: Diversity, Seasonality, and Prevalence of Borrelia burgdorferi and Anaplasma phagocytophilum.

Ixodes spp. are commonly found on dogs and cats throughout the world. In the eastern United States, 16S rDNA sequence of Ixodes scapularis, the predominant species, reveals two clades-American and Southern. To confirm the species and clades of Ixodes spp. ticks submitted from pets, we examined ticks morphologically and evaluated 16S rDNA sequence from 500 ticks submitted from 253 dogs, 99 cats, 1 rabbit, and 1 ferret from 41 states. To estimate pathogen prevalence, flaB of Borrelia burgdorferi (Bb) sensu stricto and 16S rDNA of Anaplasma phagocytophilum (Ap) were amplified and sequenced. Most Ixodes spp. from the Northeast (n = 115/115; 100%) and the Midwest (n = 77/80; 96.3%) were I. scapularis, American clade. Borrelia spp. were identified in 34 of 192 (17.8%) and Ap in 5 of 192 (2.6%) I. scapularis. Two Ixodes cookei and one Ixodes texanus were identified from Ohio, Illinois, and Michigan. In contrast, 156 of 261 (59.8%) Ixodes spp. from the Southeast were I. scapularis, American clade; 86 of 261 (33.0%) were I. scapularis, Southern clade; 9 of 261 (3.4%) were Ixodes affinis; and 10 of 261 (3.8%) were I. cookei. Southern clade was significantly more common in Florida and less common in the upper South (p < 0.0001). One I. scapularis (1/242; 0.4%) from the Southeast (Kentucky) tested positive for Bb and 6 of 242 (2.5%) were positive for Ap. In the West, most (34/44; 77.3%) Ixodes spp. were Ixodes pacificus, with Ixodes angustus (n = 6) submitted from dogs in Alaska, Washington, and Oregon and Ixodes haerlei (n = 4) preliminarily identified from a dog in Montana. Pathogens were not detected in any ticks from the West. Although I. scapularis, American clade, predominated in the Northeast and Midwest, additional Ixodes spp. were found on dogs and cats in other regions and pathogens were less commonly detected. The role of less common Ixodes spp. as disease vectors, if any, warrants continued investigation.

Recent reports of winter tick, Dermacentor albipictus, from dogs and cats in North America.

Dermacentor albipictus, a common one-host tick of large animals in North America, is most often reported from moose (Alces alces) and is rarely implicated as a parasite of cats and dogs. From 2018 to 2020, 4 dogs and 4 cats from United States and 3 dogs from Canada were infested with D. albipictus. The specimens were collected and submitted to university diagnostic specialists by veterinary clinics in Indiana, Kentucky, Minnesota, Colorado, Wyoming, Utah, Alberta, and British Columbia between the months of October to February (United States) and April to June (Canada). Six adults and five nymphal D. albipictus were collected in the United States while three adults were collected from pets in Canada, and most often a single D. albipictus was present. Identification of specimens collected in the United States were confirmed by amplification and sequencing of 16S rRNA and ITS-2 gene fragments. Rickettsia spp. were not detected in any D. albipictus collected in the United States by 17 kDa-based PCR. As tick populations continue to increase and expand in North America, correct identification of ticks collected from pets is critical to accurately track the progression and spread of ticks and tick-borne diseases.

Heartworm prevalence in dogs versus cats: Multiple diagnostic modalities provide new insights.

The cornerstones of diagnosis of heartworm (HW) in dogs are the detection of circulating antigen from adult female Dirofilaria immitis or the visualization of microfilariae in whole blood. These tests are less sensitive in cats because of the feline immune response leading to low numbers of adult worms, but heartworm antibody tests are also licensed for use in cats. HW antibodies in cats are detectable when there has at least been larval development in the tissues, but positive antibody tests cannot distinguish between current and previous larval infections; thus, cats with positive antibody test results are considered currently or previously infected with D. immitis. The aim of the present study was to use multiple HW diagnostic modalities to maximize detection of infection in dogs and cats at high risk of infection and to compare infection prevalence between these two hosts. Blood samples collected from 100 stray dogs and 100 stray cats at Florida animal shelters were tested for HW antigen (before and after heat treatment) and microfilariae; cats were also tested for HW antibody. Dogs were significantly (P = 0.0001) more likely to be diagnosed with adult HW infection (28 %; 95 % CI: 20.1-37.6%) when compared with cats (4 %; 95 % CI: 1.6-10.2%) on the basis of positive antigen and microfilariae test results. Cats with current or previous adult, immature adult, or larval HW infections comprised 19 % (95 % CI: 12.4%-27.9%) of the feline population, which was not significantly different (P = 0.1) from the prevalence of adult D. immitis infection in dogs. Testing unprotected cats for heartworm antibodies demonstrated a similar, high risk of infection to the matched unprotected dog population in Florida, which supports the use of HW preventives in cats in areas where HW transmission occurs.